Dependency on prescription opioids during and after pregnancy poses a significant health risk, both to mother and child. Newborns exposed to opioids would experience a drug withdrawal syndrome and have elevated risk of cognitive disorders. Clinical care for these exposed neonates, however, is challenged by a current fundamental gap in knowledge: lack of reliable biomarkers available to objectively assess newborn's risk from drug-exposure: it is poorly understood how pre- and postnatal opioid use affects offsprings, particularly with neurodevelopment, and no reliable biomarkers are available to objectively assess a newborn's risk from drug- exposure. We seek to advance a new assay platform to effectively monitor newborns' exposure to oxycodone (oxy). Our approach will be based on two innovative approaches: extracellular vesicles (EVs) as a biomarker and iMEX (integrated magneto-electrochemical exosome) as a sensor platform. Aim 1. We will identify EV protein signatures of high-risk oxy-exposure. We will use our rodent models to emulate in-utero and postnatal oxy-exposures. EVs from brains of oxy-exposed offspring will be collected and analyzed via quantitative proteomics to identify differentially-expressed proteins. Aim 2. We will implement the second generation iMEX with significantly expand analytical capacities. We will enhance iMEX detection sensitivity by exploring a new signal amplification (nanoplasmonics); establish a unified assay to detect both transmembrane and intravesicular markers; and construct a high-throughput detector (a 96 well-plate format). This new system will be used to screen plasma EVs from oxy-exposed animals. The success of this project will generate comprehensive protein data on brain-derived EVs under oxy-exposure and critically EVs' potential as a biomarker. Furthermore, the insights gained will set the stage to further extend HiMEX technology for clinical validation. Our long-term goal is to deliver a minimally-invasive blood test for risk assessment and timely intervention for oxy-exposure.